[Differentiation of human telomerase reverse transcriptase immortalized human marrow mesenchymal stem cell into chondrocyte].
OBJECTIVE
To establish an immortalized marrow mesenchymal stem cell line to facilitate advances in cartilage engineering research.


METHODS
Human telomerase reverse transcriptase (hTERT) cDNA was transferred into primary human marrow mesenchymal stem cells (hMSC) by retroviral vector pLEGFP-C1-hTERT. Subsequently G418 resistant cell clone was screened and expanded for further studies. hMSC biomarkers and hTERT expression were confirmed by examination. Transfected hMSC was induced to differentiate into chondrocyte using TGF-P1 and dexamethasone.


RESULTS
Up-regulated hTERT expression was detected in transfected hMSC. hMSC-hTERT cells could be induced to differentiate into chondrocyte. Higher telomerase activity in transfected cells was maintained for 50 population doublings so far. Collagen II could be detected in induced transfected hMSC by immunocytochemical and hybridization in situ.


CONCLUSIONS
Ectopic expression of hTERT can effectively immortalize hMSC in vitro. Immortalized hMSC can be induced to differentiate into chondrocyte under certain condition. It may be an ideal target of further studies in cartilage engineering.
Qi
Zhi-Ming
ZM
Department of Orthopaedics, the First Affiliated Hospital of Medical University of China, Shenyang 110001, China. qzhming@yahoo.com.cn
Lü
Gang
G
Bai
Yan-Dong
YD
Wang
Hong
H
Wang
Ling
L
D001854:Bone Marrow Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D002461:Cell Line, Transformed; D019902:Chondrocytes / Q000166:cytology*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D019098:Telomerase / Q000235:genetics* / Q000378:metabolism; D014162:Transfection
C509186:TERT protein, human; D019098:Telomerase